Journal: iScience

Article Title: Schisandrin B promotes hepatic differentiation from human umbilical cord mesenchymal stem cells

doi: 10.1016/j.isci.2024.108912

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-HIF-1a , Santa Cruz Biotechnology , Cat#sc-13515; RRID: AB_627723.

Techniques: Recombinant, CCK-8 Assay, Reverse Transcription, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Staining, Software

Journal: Acta pharmaceutica Sinica. B

Article Title: Disruption of adipocyte HIF-1 α improves atherosclerosis through the inhibition of ceramide generation.

doi: 10.1016/j.apsb.2021.10.001

Figure Lengend Snippet: Figure 1 HIF-1a deficiency in adipocytes alleviates WD-induced atherosclerosis. Eight-week-old Hif1afl/flApoEe/e and Hif1aDAdipo ApoEe/e

Article Snippet: Mouse monoclonal antibody against SMPD3 (Santa Cruz Biotechnology, Texas, USA; #sc-166637), mouse monoclonal antibody against HIF-1a (Novus, CO, USA; #NB100-105), rabbit polyclonal antibody against EIF5 (Santa Cruz Biotechnology, Texas, USA; #sc-282).

Techniques:

Journal: Acta pharmaceutica Sinica. B

Article Title: Disruption of adipocyte HIF-1 α improves atherosclerosis through the inhibition of ceramide generation.

doi: 10.1016/j.apsb.2021.10.001

Figure Lengend Snippet: Figure 2 Adipocyte-specific HIF-1a ablation decreases ceramide generation. Eight-week-old Hif1afl/flApoEe/e and Hif1aDAdipo ApoEe/e mice were fed a WD for 8 weeks. (A) PLS-DA analysis of lipid metabolites in eWAT between the Hif1afl/flApoEe/e (green circle) and Hif1aDAdipo

Article Snippet: Mouse monoclonal antibody against SMPD3 (Santa Cruz Biotechnology, Texas, USA; #sc-166637), mouse monoclonal antibody against HIF-1a (Novus, CO, USA; #NB100-105), rabbit polyclonal antibody against EIF5 (Santa Cruz Biotechnology, Texas, USA; #sc-282).

Techniques:

Journal: Acta pharmaceutica Sinica. B

Article Title: Disruption of adipocyte HIF-1 α improves atherosclerosis through the inhibition of ceramide generation.

doi: 10.1016/j.apsb.2021.10.001

Figure Lengend Snippet: Figure 3 The ceramide-generation-related gene Smpd3 is a novel target gene of adipocyte HIF-1a. (A) The mRNA levels of genes related to ceramide metabolism, including synthesis, transportation and elimination in the eWAT of the AdHif1aLSL/LSL and Hif1aþ/þ mice (n Z 6 per group). Two-tailed Student’s t-test: **P < 0.01 compared to the Hif1aþ/þ mice. (B) The protein levels of HIF-1a and SMPD3 in the eWAT of the AdHif1aLSL/LSL and Hif1aþ/þ mice (n Z 3 per group). (C) The mRNA levels of genes related to ceramide metabolism, including synthesis, transportation and elimination in the eWAT of the Hif1afl/flApoEe/e and Hif1aDAdipo ApoEe/e mice fed a WD for 8 weeks (n Z 6 per group). Two-tailed Student’s t-test: **P < 0.01 compared to the Hif1afl/flApoEe/e mice. (D) The protein levels of HIF-1a and SMPD3 in the eWAT of the Hif1afl/flApoEe/e and Hif1aDAdipo ApoEe/e mice mice fed a WD for 8 weeks (n Z 3 per group). (E) Schematic diagram of the mouse Smpd3 promoter illustrating the predicted HRE in the regulatory region. The upstream regions are numbered in relation to the transcription initiation site, which is designated þ1. (F) ChIP assays of 3T3-L1 adipocytes treated with CoCl2 (200 mmol/L) for 12 h (n Z 4 per group). One-way ANOVA with Tukey’s post hoc test: **P < 0.01 compared to the IgG-Ab group. (G) Luciferase reporter gene assay of Smpd3 promoter activity (n Z 4 per group). One-way ANOVA with Dunnett’s T3 post hoc test: **P < 0.01 compared to the Smpd3 promoter plasmid with vector group; ##P < 0.01 compared to the Smpd3 promoter plasmid with HIF-1a TM group.

Article Snippet: Mouse monoclonal antibody against SMPD3 (Santa Cruz Biotechnology, Texas, USA; #sc-166637), mouse monoclonal antibody against HIF-1a (Novus, CO, USA; #NB100-105), rabbit polyclonal antibody against EIF5 (Santa Cruz Biotechnology, Texas, USA; #sc-282).

Techniques: Two Tailed Test, Luciferase, Reporter Gene Assay, Activity Assay, Plasmid Preparation

Journal: Acta pharmaceutica Sinica. B

Article Title: Disruption of adipocyte HIF-1 α improves atherosclerosis through the inhibition of ceramide generation.

doi: 10.1016/j.apsb.2021.10.001

Figure Lengend Snippet: Figure 6 Administration of ceramide reverses the protective effects of adipose HIF-1a deficiency on atherosclerosis through increased inflammation and cholesterol. Eight-week-old Hif1afl/flApoEe/e and Hif1aDAdipo ApoEe/e mice were injected with ceramide C16:0 or vehicle and were fed a WD for 6 weeks. (A) Schematic diagram of the mouse model illustrating the concept of mouse injection with ceramide C16:0 or vehicle. (B, C) Quantitation of ceramide concentrations in plasma (B) and eWAT (C), n Z 6 per group. (D, E) The levels of plasma total cholesterol (D) and triglyceride (E), n Z 6e7 per group. (F) FPLC analysis of plasma cholesterol levels in mice. (G) Left, representative oil red O staining of cross- sections of aortic roots. Scale bar, 500 mm. Right, quantification of aortic root lesion areas. n Z 6e7 per group. (H) Immunofluorescence stain- ing of atherosclerotic lesions with MAC-2 antibody and calculated MAC-2 positive areas in the plaques (n Z 3 per group). Scale bar, 500 mm. All data are presented as the mean SEM. One-way ANOVA with Tukey’s post hoc test (B, D, E, H) and KruskaleWallis test (C and G): *P < 0.05, **P < 0.01 compared to the Hif1afl/flApoEe/e þ vehicle group; #P < 0.05, ##P < 0.01 compared to the Hif1aDAdipo ApoEe/e

Article Snippet: Mouse monoclonal antibody against SMPD3 (Santa Cruz Biotechnology, Texas, USA; #sc-166637), mouse monoclonal antibody against HIF-1a (Novus, CO, USA; #NB100-105), rabbit polyclonal antibody against EIF5 (Santa Cruz Biotechnology, Texas, USA; #sc-282).

Techniques: Injection, Quantitation Assay, Clinical Proteomics, Staining

Journal: Acta pharmaceutica Sinica. B

Article Title: Disruption of adipocyte HIF-1 α improves atherosclerosis through the inhibition of ceramide generation.

doi: 10.1016/j.apsb.2021.10.001

Figure Lengend Snippet: Figure 7 Inhibition of HIF-1a by PX-478 improves atherosclerosis through the SMPD3eceramide axis. Eight-week-old ApoEe/e mice were administered with PX-478 (5 mg/kg) or vehicle every other day and fed a WD for 8 weeks. (A) Schematic diagram of the mouse model illustrating the concept of mouse injection with PX-478. (B) Left, representative oil red O staining of cross-sections of aortic roots. Scale bar, 500 mm. Right, quantification of aortic root lesion areas. n Z 7 per group. (C, D) Quantitation of ceramide concentrations in plasma (C) and eWAT (D), n Z 6 per group. (E, F) The levels of plasma total cholesterol (E) and triglyceride (F), n Z 7 per group. (G) FPLC analysis of plasma cholesterol levels in mice. (H) Immunofluorescence staining of atherosclerotic lesions with MAC-2 antibody and calculated MAC-2 positive areas in the plaques (n Z 3 per group). Scale bar, 500 mm. All data are presented as the mean SEM. ManneWhitney U test (B, D), Two-tailed Student’s t-test (C, E, F, H): *P < 0.05, **P < 0.01 compared to the vehicle group. See also Fig. S5.

Article Snippet: Mouse monoclonal antibody against SMPD3 (Santa Cruz Biotechnology, Texas, USA; #sc-166637), mouse monoclonal antibody against HIF-1a (Novus, CO, USA; #NB100-105), rabbit polyclonal antibody against EIF5 (Santa Cruz Biotechnology, Texas, USA; #sc-282).

Techniques: Inhibition, Injection, Staining, Quantitation Assay, Clinical Proteomics, Two Tailed Test